中华皮肤科杂志 ›› 2002, Vol. 35 ›› Issue (5): 340-342.

• 论著 • 上一篇    下一篇

微孔板双探针杂交法快速鉴别都柏林念珠菌和白念珠菌

朱利平1, 章强强2, 石尧忠1, 李莉2, 王家俊2, 翁心华1   

  1. 1. 复旦大学附属华山医院感染科, 上海200040;
    2. 复旦大学附属华山医院皮肤科, 上海200040
  • 收稿日期:2002-04-15 出版日期:2002-10-15 发布日期:2002-10-15
  • 基金资助:
    2001年中华医学会皮肤科分会科研基金

Rapid Identification of Candida albicans and Candida dubliniensis Isolates With Two Specific Probes Simultaneously

ZHU Liping1, ZHANG Qiangqiang2, SHI Yaozhong1, LI Li2, WANG Jiajun2, WENG Xinhua1   

  1. Department of Infectious Diseases and Dermatology, Huashan Hospital, Fudan University, Shanghai 200040, China
  • Received:2002-04-15 Online:2002-10-15 Published:2002-10-15

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摘要： 目的 建立微孔板双探针杂交法,快速鉴别都柏林念珠菌和白念珠菌,并应用于临床分离株的快速鉴定。方法 利用真菌保守区核糖体基因和可变区内转录间隔区基因为靶序列,应用生物素标记的通用引物进行念珠菌DNA的PCR扩增,并将该PCR产物分别与固定在微孔板上的都柏林念珠菌和白念珠菌特异性探针杂交,经酶联显色反应测其A值。结果 能特异地鉴别白念珠菌和都柏林念珠菌标准菌株。对108株常规培养方法从临床标本分离的白念珠菌检测,结果显示106株菌株仅白念珠菌探针检测阳性,另2株菌株仅都柏林念珠菌探针阳性。结论 微孔板双探针杂交法能快速、特异地鉴别都柏林念珠菌和白念珠菌。

关键词: 关键词念珠菌,都柏林, 念珠菌,白色, 核酸杂交法, DNA探针

Abstract: Objective To develop a microtitration plate enzyme immunoassay to differentiate Candida albicans from Candida dubliniensis isolates using two specific probes simultaneously.Methods The fun-gus-specific universal primers derived from the internal transcribed spacer region of fungal rDNA were labeled with biotin,while the C.albicans or C.dubliniensis specific capture probes were coated on the microplates.Genomic DNA purified from the two species was amplified by PCR.The biotinylated products were captured by the probes coated on the microplates.The A405 value was finally determined by the colorimetric assay.Results The two species of Candida could be detected specifically.Out of 108 clinical isolates originally identified as C.albicans on the basis of germtube formation,two isolates were positive for C.dubliniensis and negative for C.albicans.The other106isolates were positive for C.albicans and negative for C.dubliniensis.Conclusions Two-specific-probe hybridization method is rapid and reliable for differentiating C.albicans from C.dubliniensis.

Key words: Candida albicans, Candida dubliniensis, Nucleic acid hybridization, DNA probe